We offer a broad range of real time PCR/ RT-PCR reagents to meet various experimental needs.Our qPCR portfolio comprises both SYBR Green and probe-based qPCR, Singleplex and multiplex qPCR, and One-step and two-step qRT-PCR. How to choose a suitable real time PCR/ RT-PCR reagent solution is depending on the features of each category.
SYBR Green or probe-based qPCR?
(1)SYBR Green qPCR features: cost-effective, only a single amplicon can be detected, inaccurate quantification.
(2)Probe-based qPCR features: increased cost due to the design of probes and primers, accurate quantification with high specificity, multiplex reactions permitted.
(3)Selection recommendations: for low-throughput, singleplex experiments, SYBR Green method is preferable. For high-throughput or multiplex experiments, probe-based method with high specificity is desirable because the initial cost can be spread over many assays for high-throughput experiments and specificity is required for multiplex assays.
Singleplex or multiplex qPCR?
(1)Singleplex qPCR features: easier to design, decreased instrument cost, increased sample material requirements and time consumption.
(2)Multiplex qPCR features: decreased sample material cost, internal control permitted, suitable for high-throughput and high-volume analyses, decreased risk for pipetting errors, a broader range of applications including SNP genotyping and pathogen detection, multiple optimization rounds required.
(3)Selection recommendations: for experiments with limited amount of starting materials, requiring an internal control or high throughput analysis, multiplex qPCR is recommended, otherwise singleplex qPCR is sufficient.
One-step or two-step qRT-PCR?
(1)One-step qRT-PCR features: easier to set up with less handling time, reduced contamination, no stock of cDNA for future use, inability to optimize reverse transcription step.
(2)Two-step qRT-PCR features: allowing for stocking of cDNA, flexible to optimize and control, with stock of cDNA for future use, more time consuming.
(3)Selection recommendations: for applications requiring high-speed and high-throughput amplification of many samples with few targets, one-step qRT-PCR is recommended. For applications with limited sample input or requiring cDNA stocks, two-step qRT-PCR may be more suitable.
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